Abstracting and Indexing

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Annotation of Ribosomal Protein Mass Peaks in MALDI-TOF Mass Spectra of Bacterial Species and Their Phylogenetic Significance

Author(s): Wenfa Ng

Although MALDI-TOF mass spectrometry based microbial identification has achieved a level of accuracy that facilitate its use in classifying microbes to the species and strain level, questions remain on the identities of the mass peaks profiled from individual microbial species. Specifically, in the popular approach of comparing the mass spectrum of known and unknown microbes for identification purposes, the identities of the mass peaks were not taken into consideration. This study sought to determine if ribosomal proteins could account for some of the mass peaks profiled in MALDI-TOF mass spectra of different bacterial species. Using calculated molecular mass of ribosomal proteins for annotating mass peaks in bacterial species’ MALDITOF mass spectra downloaded from the SpectraBank database, this study revealed that ribosomal proteins could account for the low molecular weight mass peaks of <10000 Da. However, contrary to published reports, ribosomal proteins could not account for most of the mass peaks profiled. In particular, the data revealed that between 1 and 6 ribosomal protein mass peaks could be annotated in each mass spectrum. Annotated ribosomal proteins were S16, S17, S18, S20 and S21 from the small ribosome subunit, and L27, L28, L29, L30, L31, L31 Type B, L32, L33, L34, L35 and L36 from the large ribosome subunit. The ribosomal proteins with the most number of mass peak annotations were L36 and L29, with L34, L33, and L31 completing the list of ribosomal proteins with large number of annotations. Given the highly conserved nature of most ribosomal proteins, possible phylogenetic significance of the annotated ribosomal proteins were investigated through reconstruction of maximum likelihood phylogenetic trees. Results revealed that except for ribosomal protein L34, L31, L36 and S18, all annotated ribosomal proteins hold phylogenetic significance under the criteria of recapitulation of phylogenetic cluster groups present in the phylogeny of 16S rRNA. Phylogenetic significance of the annotated ribosomal proteins was further verified by the phylogenetic tree constructed based on the concatenated amino acid sequence of L29, S16, S20, S17, L27 and L35. Finally, analysis of the structure of the annotated ribosomal proteins did not reveal a high conservation of structure of the ribosomal proteins. Collectively, small low molecular weight (<10000 Da) ribosomal proteins could annotate some of the mass peaks in MALDI-TOF mass spectra of various bacterial species, and most of the ribosomal proteins hold phylogenetic significance. However, structural analysis did not identify a conserved structure for the annotated ribosomal proteins. Annotation of ribosomal protein mass peaks in MALDI-TOF mass spectra highlighted the deep biological basis inherent in the mass spectrometry-based microbial identification method.

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